Insig1

Antibodies to ACC1-pS79, ACC1, FASN, SCD1, Tubulin, TBP, GAPDH, AKT-pS473, AKT, GSK3β-pS21/9, GSK3β, TSC2-pT1462, TSC2, mTOR-pS2448, mTOR, S6K-pT389, S6K, S6-pS240/244, S6, 4E-BP1-pS65, 4E-BP1, β-actin, LRP6-pS1490, LRP6, TCF7L2, β-catenin, β-catenin-pS33/37/T41, IGF1R, AKT-pT308, and Lamin B were purchased from Cell Signaling. Antibodies to apoB, SCAP, Insig1, Insig2, Sp5, Sp1 were purchased from Santa Cruz Biotech (Santa Cruz, Ca, USA). Antibodies to MTP, SREBP1, SREBP2, IRS1 were purchased from BD Biosciences. Antibodies to ACAT2 and IGF1 were purchased from Novus. Antibodies to ELOVL6 (Thermo Scientific), DGAT1 (Bio Vision), GPAT1 (GeneTex), LXRα (Abcam), HMGCR (Upstate), and IRS1-pY612 (Invitrogen) were used for western blotting.

Knock-down of selected genes was achieved by transfecting THP-1 macrophages using lipofectamine RNAiMax (Invitrogen) according to manufacturer’s instructions. Cells were transfected with 50 nM control scrambled siRNA (Invitrogen) or siRNA targeting genes of interest, FADS1, INSIG1, and LDLR (Santa Cruz Biotechnology).